title: A new method for producing transgenic birds via direct in vivo transfection of primordial germ cells.
abstract: Traditional methods of avian transgenesis involve complex manipulations involving either retroviral infection of blastoderms or the ex vivo manipulation of primordial germ cells (PGCs) followed by injection of the cells back into a recipient embryo. Unlike in mammalian systems, avian embryonic PGCs undergo a migration through the vasculature on their path to the gonad where they become the sperm or ova producing cells. In a development which simplifies the procedure of creating transgenic chickens we have shown that PGCs are directly transfectable in vivo using commonly available transfection reagents. We used Lipofectamine 2000 complexed with Tol2 transposon and transposase plasmids to stably transform PGCs in vivo generating transgenic offspring that express a reporter gene carried in the transposon. The process has been shown to be highly effective and as robust as the other methods used to create germ-line transgenic chickens while substantially reducing time, infrastructure and reagents required. The method described here defines a simple direct approach for transgenic chicken production, allowing researchers without extensive PGC culturing facilities or skills with retroviruses to produce transgenic chickens for wide-ranging applications in research, biotechnology and agriculture.
authors.affiliation: CSIRO Biosecurity Flagship, Australian Animal Health Laboratory, Geelong, VIC, 3220, Australia.
authors.forename: Scott G
authors.initials: SG
authors.lastname: Tyack
authors.forename: Kristie A
authors.initials: KA
authors.lastname: Jenkins
authors.forename: Terri E
authors.initials: TE
authors.lastname: O'Neil
authors.forename: Terry G
authors.initials: TG
authors.lastname: Wise
authors.forename: Kirsten R
authors.initials: KR
authors.lastname: Morris
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Bruce
authors.forename: Scott
authors.initials: S
authors.lastname: McLeod
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Wade
authors.forename: James
authors.initials: J
authors.lastname: McKay
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Moore
authors.forename: Karel A
authors.initials: KA
authors.lastname: Schat
authors.forename: John W
authors.initials: JW
authors.lastname: Lowenthal
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Doran
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002645:Chickens / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics*; D018014:Gene Transfer Techniques; D005854:Germ Cells; D008055:Lipids / Q000235:genetics; D010957:Plasmids; D014162:Transfection / Q000379:methods
chemical_list: D004251:DNA Transposable Elements; D008055:Lipids; C086724:Lipofectamine